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Page 1: K. Mikula, M. Smíšek - math.sk · Filtering, cell center detection and cell segmentation by geometrical partial differential equations K. Mikula, M. Smíšek Automatic image analysis

Filtering, cell center detection and cell segmentation by

geometrical partial differential equations

K. Mikula, M. Smíšek

Automatic image analysis

By image analysis we mean extraction of cell centers and

shape of each cell from microscopy image.

By automatic image analysis we mean the image analysis

performed by computer algorithms, without human inputs.

By image processing chain we mean interconnected chain of

processes performing automatic image analysis. We identify

these three image processing chain processes:[0]

• Filtering of the input image (using GMCF)

• Cell center identification (using LSCD)

• Segmentation of cell shapes (using GSUBSURF)

We also propose a way to detect mitosis in given data.

Input data

The input data tested were in two dimensions + time, i.e. 2D

video. Data contains cell membrane images of drosophila and

zebrafish, both in phase of morphogenesis.

References

[0] Bourgine, P., Čunderlík, R., Drblíková-Stašová, O., Mikula, K., Peyriéras,

N., Remešíková, M., Rizzi, B., Sarti, A.: 4D embryogenesis image analysis

using PDE methods of image processing. (to appear in) Kybernetika, Vol. 46,

No. 2 (2010)

[1] Chen, Y., Vemuri, B. C., Wang, L.: Image denoising and segmentation via

nonlinear diffusion. Computers and Mathematics with Applications 39 (2000),

131–149.

[2] Frolkovič, P., Mikula, K., Peyrieras, N., Sarti, A.: A counting number of

cells and cell segmentation using advection-diffusion equations. Kybernetika

43 (6) (2007), 817–829.

[3] Sarti, A., Maladi, R., Sethian, J. A.: Subjective surfaces: A method for

completing missing boundaries. Proceedings of the National Academy of

Sciences of the United States of America 12 (97) (2000), 6258–6263.

[4] Mikula, K., Peyrieras, N., Remesikova, M., Sarti, A.: 3D embryogenesis

image segmentation by the generalized subjective surface method using the

finite volume technique. Finite Volumes for Complex Applications V:

Problems and Perspectives (Eds. R.Eymard, J.M.Herard), ISTE and WILEY,

London, 2008, pp. 585-592)

Input data – membrane images. Left – zebrafish, right – drosophila.

1. Filtering by GMCF[1]

GMCF = Geodesic Mean Curvature Flow

• removes noise structures

• preserves edges

GMCF equation:

0.

u

uuGguut

Function g with its argument is called edge detection

function and has values:

• ≈ 0 if given pixel belongs to edge

• ≈ 1 otherwise

We use this edge detector equation: (with s = )

2. Cell center identification by LSCD[2]

Goal of this step is obtaining list of approximate cell centers.

LSCD = Level Set Center Detection

LSCD equation:

for some positive constants δ and μ.

• Small contours (those of noise artifacts) implode rapidly fast

• Larger (those of cell structures) are observable for a longer

period of time

• Taking local maximae of these images gives us geometric

centers of cells

kV

0.

u

uuuut

3D plot of intensity function after LSCD. Local maximae represent

approximate cell centers pretty well. – these are plotted as red dots in the

image below.Left – zebrafish, right – drosophila.

Mitosis detection

The goal is to find splitting of two cells in space and time to

detect mitosis. We consider 2D + time video as a 3D image

and try to find trousers-like shapes – again using GSUBSURF

as segmentation algorithm. Mitosis happens, where two ‘leg-

parts’ split.

0,1

1)(

2

k

kssg

uG

Demonstration of edge detection function. Low intensity is white, high

intensity is black. Left – zebrafish, right – drosophila.

Input data shown as 3D plot of intensity function. Left – zebrafish, right –

drosophila.

• All contours advect in the direction of inner normal, and the

speed V of this advection is also a function of their curvature k:

Data filtered by GMCF. Left – zebrafish, right – drosophila.

GMCF filtered data as 3D plot. Left – zebrafish, right – drosophila.

Top – initial segmentation contour, bottom – final segmentation contour.

Left – zebrafish, right – drosophila.

For better illustration of data properties, we are working with

images after histogram equalization.

We thank to N.Peyriéras (CNRS, Gif sur Ivette) and F.Graner

(Institut Curie, Paris) for zebra-fish and drosophila testing data.

We also thank to Francois Graner for bringing our attention to

trousers-like 2D + time image segmentation.

3. Segmentation by GSUBSURF[3],[4]

Goal of segmentation is to take a cell center and say, what

is the area covered by this cell.

GSUBSURF = Generalized SUBjective SURFace

GSUBSURF equation:

0..

ugw

u

uugwu condift

Identified cell centers. Red dots are approximate cell centers – they are the

local maximae of 3D plots in the previous figure. Left – zebrafish, right –

drosophila.

Four drosophila cells and their segmentation considering time. Bottom –

cell at time t = 35, top – cell at time t = 55, center – what happens ‘in

between’.

We observe first and second cell forming a trousers-like shape in space

and time, which means cell division. The other two do not divide in this

period.

Same cells as above, now shown in correct positions in image at time

t = 35.

Drosophila segmentation. Top – initial segmentation function,

bottom – final segmentation for cells marked below.

Contact: [email protected], [email protected]

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